流式细胞仪在检测淋巴结树突状细胞中的应用

Value of flow cytometry in monitoring lymph node dendritic cells

目的利用流式细胞术多参数分析的特点对淋巴结树突状细胞含量准确计数。方法手术取宫颈癌淋巴转移和非淋巴转移患者淋巴结组织。将组织用机械法制备成单个细胞悬液,调整细胞浓度为5×106个细胞/ml。每例标本分两管,一管只加树突状细胞(dendritic cell,DC)标记物CD83抗体,另一管加CD83的同时再加白细胞表面共同抗原CD45RO,用流式细胞仪分别检测两管CD83阳性百分率。结果淋巴结转移组中双标记组和单标记组间CD83的表达差异显著;非淋巴结转移组中双标记组和单标记组间CD83的表达差异显著;淋巴结转移组与非淋巴结转移组在单标记和双标记间CD83表达率也有显著差异。结论肿瘤的转移与周围淋巴结树突状细胞减少有关。对于含量很少的树突状细胞像在淋巴结这样成分较复杂组织中计数,采用双参数分析是必要的。

Objective To count accurately the dendritic cells of lymph nodes by multiparameter flow cytometry. Methods The metastasis and non-metastasis nodes were obtained by endometrial carcinoma surgery, homogenizated to single-cell suspension with the concentration of 5 × 10^6/ml. Every case was divided into two tubes： one was added CD83 antibody only, the other was added both leukocyte common antigen CD45RO and CD83 antibody. The positive rates of CD83 antibody in both tubes were monitored by flow cytometry. Results CD83 expression was significantly different between metastasis group and non-metastasis group, and also different between double-labeling group and single-labeling group. Conclusion The tumor metastasis is concemed with the decrease of the lymph node dendritic cells. The method labeled with leukocyte common antigen CD45RO and CD83 antibody could avoid the interference and enhance the accuracy. Meanwhile, it is necessary to count the very small dendritic cells by multiparameter flow cytometry.